Research and Development Center, Otsuka Pharmaceutical Factory, Inc., Naruto, Japan.
Otsuka America Pharmaceutical, Inc. (OAPI), Schaumburg, Illinois.
Xenotransplantation. 2019 May;26(3):e12501. doi: 10.1111/xen.12501. Epub 2019 Feb 15.
Due to recent advances in research on mesenchymal stem cells (MSCs), MSCs are expected to be used in various clinical applications. However, securing adequate cadaveric donors and safety of living donors are major issues. To solve such issues, we have examined to develop clinical grade neonatal porcine bone marrow-derived MSCs (npBM-MSCs). Clinical grade neonatal porcine bone marrow cells were collected, frozen, and sent to our laboratory by air. The npBM-MSCs were isolated from thawed bone marrow cells, then frozen. The thawed npBM-MSCs were examined for CD markers and differentiated into chondrocytes, osteocytes, and adipocytes. They were compared with human bone marrow-derived MSCs (hBM-MSCs) for growth rate and size. To assess the robustness of proliferation, we compared culture medium with or without gelatin. The npBM-MSCs expressed positive MSC markers CD29, CD44, and CD90 and were differentiated into chondrocytes, osteocytes, and adipocytes. The doubling time of npBM-MSCs was significantly shorter than that of hBM-MSCs (17.3 ± 0.8 vs 62.0 ± 19.6 hours, P < 0.01). The size of npBM-MSCs was also significantly smaller than that of hBM-MSCs (13.1 ± 0.3 vs 17.5 ± 0.4 μm, P < 0.001). The npBM-MSCs showed similar proliferation characters irrespective of with or without gelatin coating. The npBM-MSCs secreted VEGF-A, VEGF-C, and TGF-β1. We have established npBM-MSCs which show super-rapid growth, small size, and robust proliferation profile. The np-MSCs might be able to solve the donor issues for MSC therapy.
由于间充质干细胞 (MSCs) 研究的最新进展，预计 MSCs 将在各种临床应用中得到应用。然而，确保充足的尸体供体和活体供体的安全性是主要问题。为了解决这些问题，我们已经研究开发了临床级新生猪骨髓来源的间充质干细胞 (npBM-MSCs)。收集、冷冻并通过空运将临床级新生猪骨髓细胞发送到我们的实验室。从解冻的骨髓细胞中分离 npBM-MSCs，然后进行冷冻。解冻后的 npBM-MSCs 检查了 CD 标志物，并分化为软骨细胞、成骨细胞和脂肪细胞。将其与人类骨髓来源的间充质干细胞 (hBM-MSCs) 进行比较，以评估其生长速度和大小。为了评估增殖的稳健性，我们比较了有或没有明胶的培养基。npBM-MSCs 表达 MSC 标志物 CD29、CD44 和 CD90，并分化为软骨细胞、成骨细胞和脂肪细胞。npBM-MSCs 的倍增时间明显短于 hBM-MSCs（17.3 ± 0.8 小时对 62.0 ± 19.6 小时，P < 0.01）。npBM-MSCs 的大小也明显小于 hBM-MSCs（13.1 ± 0.3 微米对 17.5 ± 0.4 微米，P < 0.001）。无论是否使用明胶涂层，npBM-MSCs 的增殖特性均相似。npBM-MSCs 分泌 VEGF-A、VEGF-C 和 TGF-β1。我们已经建立了 npBM-MSCs，它们表现出超快的生长速度、较小的大小和稳健的增殖特征。np-MSCs 可能能够解决 MSC 治疗的供体问题。
